Ferritin reduction is essential for cerebral ischemia-induced hippocampal neuronal death through p53/SLC7A11-mediated ferroptosis.

Cerebral ischemia is the most common cause of hippocampal neuronal death and the most prevalent cause of stroke with high mortality rate. Ferroptosis has been suggested to affect the role of hippocampal neurons. This study explores the influence of lentivirus infection-induced ferritin overexpression in hippocampal neuronal injury and death through simulations in August Copenhagen Irish rat models. Twenty-four-hour cerebral ischemia-reperfusion injury was induced in the rats after 90-min middle cerebral artery occlusion (MCAO). Ferritin overexpression was induced through lentivirus infection. The Morris Water Maze (MWM) test and tau hyperphosphorylation test were performed on hippocampal neurons to establish a MCAO model. The effect of ferritin overexpression on hippocampal neuronal death was evaluated using hematoxylin-eosin staining and annexin V/propidium iodide flow cytometry. The MWM test revealed that MCAO modeling decreased the cognitive and locomotor capacity of the rats, whereas ferritin overexpression partially reversed the effect of MCAO. In addition, the hyperphosphorylation of tau caused by MCAO was reduced by ferritin. Pathogenic changes, impaired viability, increased apoptosis, and elevated caspase-9 cleavage in hippocampal neurons were clearly recovered by ferritin. Moreover, robust reactive oxygen species production and glutathione consumption, which was induced by MCAO modeling, were ameliorated by ferritin. Furthermore, two key modulators of ferroptosis, p53 and SLC7A11, were demonstrated to be upregulated by MCAO modeling and downregulated by ferritin. Ferritin reduction is essential for cerebral ischemia-induced hippocampal neuronal ferroptosis mediated via p53 and SLC7A11.

Catheter dwell time and risk of catheter failure in adult patients with peripheral venous catheters.

AIMS AND OBJECTIVES: To explore whether the risk of peripheral venous catheters failure remained constant throughout catheter use in adult patients. BACKGROUND: Peripheral venous catheters, widely used in adult patients, may have a critical threshold dwell time associated with increased risk of catheter failure. DESIGN: Prospective, observational study. We have complied with the STROBE checklist of items. METHODS: This study was conducted from July-October 2018 in Hunan, China. Data on patient factors, catheter factors and catheter failure events were collected. Poisson regression was used to assess the effect of catheter dwell time on catheter failure while adjusting for other variables. RESULTS: A total of 1,477 patients were included in the analysis. There were 854 cases (57.8%) of catheter failure. The median dwell time to catheter failure was 52 hr (interquartile range: 36-73 hr). The incidence rate of catheter failure significantly increased by 1.1%/h in the first 38 hr after catheter insertion. From 39-149 hr, the incidence rate significantly decreased, and at >149 hr, there was no significant change in the incidence rate. Meanwhile, factors such as vascular quality and infused drugs showed having an impact on catheter failure events. CONCLUSIONS: The risk of catheter failure may not remain constant throughout the dwell time. The results suggest that nurses should assess the insertion site frequently in the first 38 hr. RELEVANCE TO CLINICAL PRACTICE: The significant increase in the risk of catheter failure per hour may warrant close and frequent inspection of insertion site during the first 38 hr.

Nano-LC-ESI MS/MS analysis of proteins in dried sea dragon Solenognathus hardwickii and bioinformatic analysis of its protein expression profiling / 中国天然药物

The sea dragon Solenognathus hardwickii has long been used as a traditional Chinese medicine for the treatment of various diseases, such as male impotency. To gain a comprehensive insight into the protein components of the sea dragon, shotgun proteomic analysis of its protein expression profiling was conducted in the present study. Proteins were extracted from dried sea dragon using a trichloroacetic acid/acetone precipitation method and then separated by SDS-PAGE. The protein bands were cut from the gel and digested by trypsin to generate peptide mixture. The peptide fragments were then analyzed using nano liquid chromatography tandem mass spectrometry (nano-LC-ESI MS/MS). 810 proteins and 1 577 peptides were identified in the dried sea dragon. The identified proteins exhibited molecular weight values ranging from 1 900 to 3 516 900 Da and pI values from 3.8 to 12.18. Bioinformatic analysis was conducted using the DAVID Bioinformatics Resources 6.7 Gene Ontology (GO) analysis tool to explore possible functions of the identified proteins. Ascribed functions of the proteins mainly included intracellular non-membrane-bound organelle, non-membrane-bounded organelle, cytoskeleton, structural molecule activity, calcium ion binding and etc. Furthermore, possible signal networks of the identified proteins were predicted using STRING (Search Tool for the Retrieval of Interacting Genes) database. Ribosomal protein synthesis was found to play an important role in the signal network. The results of this study, to best of our knowledge, were the first to provide a reference proteome profile for the sea dragon, and would aid in the understanding of the expression and functions of the identified proteins.

Agglutinin isolated from Arisema heterophyllum Blume induces apoptosis and autophagy in A549 cells through inhibiting PI3K/Akt pathway and inducing ER stress / 中国天然药物

Arisaema heterophyllum Blume is one of the three medicinal plants known as traditional Chinese medicine Rhizoma Arisaematis (RA). RA has been popularly used to treat patients with convulsions, inflammation, and cancer for a long time. However, the underlying mechanisms for RA effects are still unclear. The present study was designed to determine the cytotoxicity of agglutinin isolated from Arisema heterophyllum Blume (AHA) and explore the possible mechanisms in human non-small-cell lung cancer A549 cells. AHA with purity up to 95% was isolated and purified from Arisaema heterophyllum Blume using hydrophobic interaction chromatography. AHA dose-dependently inhibited the proliferation of A549 cells and induced G phase cell cycle arrest. AHA induced apoptosis by up-regulating pro-apoptotic Bax, decreasing anti-apoptotic Bcl-2, and activating caspase-9 and caspase-3. In A549 cells treated with AHA, the PI3K/Akt pathway was inhibited. Furthermore, AHA induced increase in the levels of ER stress markers such as phosphorylated eukaryotic initiation factor 2α (p-eIF2α), C/EBP-homologous protein (CHOP), inositol-requiring enzyme 1α (IRE1α), and phosphorylated c-Jun NH-terminal kinase (p-JNK). AHA also induced autophagy in A549 cells. Staining of acidic vesicular organelles (AVOs) and increase in the levels of LC3II and ATG7 were observed in AHA-treated cells. These findings suggested that AHA might be one of the active components with anti-cancer effects in Arisaema heterophyllum Blume. In conclusion, cytotoxicity of AHA on cancer cells might be related to its effects on apoptosis and autophagy through inhibition of PI3K/Akt pathway and induction of ER stress.

Proteomic and bioinformatic analyses of possible target-related proteins of gambogic acid in human breast carcinoma MDA-MB-231 cells / 中国天然药物

Gambogic acid (GA) is an anticancer agent in phase ‖b clinical trial in China but its mechanism of action has not been fully clarified. The present study was designed to search the possible target-related proteins of GA in cancer cells using proteomic method and establish possible network using bioinformatic analysis. Cytotoxicity and anti-migration effects of GA in MDA-MB-231 cells were checked using MTT assay, flow cytometry, wound migration assay, and chamber migration assay. Possible target-related proteins of GA at early (3 h) and late stage (24 h) of treatment were searched using a proteomic technology, two-dimensional electrophoresis (2-DE). The possible network of GA was established using bioinformatic analysis. The intracellular expression levels of vimentin, keratin 18, and calumenin were determined using Western blotting. GA inhibited cell proliferation and induced cell cycle arrest at G2/M phase and apoptosis in MDA-MB-231 cells. Additionally, GA exhibited anti-migration effects at non-toxic doses. In 2-DE analysis, totally 23 possible GA targeted proteins were found, including those with functions in cytoskeleton and transport, regulation of redox state, metabolism, ubiquitin-proteasome system, transcription and translation, protein transport and modification, and cytokine. Network analysis of these proteins suggested that cytoskeleton-related proteins might play important roles in the effects of GA. Results of Western blotting confirmed the cleavage of vimentin, increase in keratin 18, and decrease in calumenin levels in GA-treated cells. In summary, GA is a multi-target compound and its anti-cancer effects may be based on several target-related proteins such as cytoskeleton-related proteins.

[Study on the choice of functional monomer before preparation of myclobutanil molecularly imprinted polymer].

To prepare myclobutanil molecularly imprinted polymer, a method was established for the choice of the appropriate functional monomer and its dosage. UV spectra was applied to study the combination form, the effect intensity, the optimal concentration ratio and the numbers of binding sites between myclobutanil and methyl acrylic acid (MAA) or acrylamide (AM) functional monomer. The results showed that hydrogen-bonding interaction could be formed between myclobutanil and methyl acrylic acid (MAA) or acrylamide (AM) functional monomer. The pi electron of the triazole ring conjugated double bond in my clobutanil could transit to pi* conjugate antibonding orbital when it absorbed energy. The formation of hydrogen bond could make pi-->pi* absorption band transit. Maximum absorption wavelength produced red shift with the increase in the functional monomer concentration in the system. The research revealed that the optimal concentration ratios between myclobutanil and the two monomers were c(M):c(MAA) = 1:4, c(M):c(AM) = 1:2. Myclobutanil and the both the functional monomers had the bonding ability, and strong bonding force. The prepared molecularly imprinted polymer using AM as a functional monomer had better stability and specificity of recognition for myclobutanil.

[Quality evaluation of artificial musk based on its inhibitory effect on cyclooxygenase-2].

The inhibitory ratio (1%) of artificial musk on cyclooxygenase-2 (COX-2) was determined by enzyme immunoassay (EIA). The dose-effect relationship between concentrations of artificial musk and 1% was established. It was found that artificial musk had obvious inhibitory action on COX-2. The concentration for 50% of maximum inhibitory effect (IC50) was about 2.26 mg x mL(-1). There was a good relationship between the logarithm concentrations of artificial musk and 1% when the concentrations of artificial musk ranged from 0.31-20.0 mg x mL(-1). The results indicated that this EIA method could be applied to evaluate the anti-inflammatory activity of artificial musk quickly, conveniently, sensitively and exactly. This paper provided a novel method and foundational research for the bioassay of artificial musk.

[Microcalorimetric study on the metabolism of Staphylococcus aureus affected by qingkailing injection].

This paper is to report the investigation on the metabolic behavior of Staphylococcus aureus (S. aureus) after given Qingkailing injection, and with the aim of seeking for a new quality control method based on biological assessment. The growth thermogenic curves of S. aureus were determined by microcalorimetry and analyzed by computer. The results showed that in the concentration range of (0-5.00%), the growth thermogenic curves of S. aureus were declined and removed back with increasing dosage of Qingkailing injection; the main parameters (T1, T2, k1, P1, P2 and I) and the dosage of Qingkailing injection have good correlation. The 50% inhibiting dosage is 3.26 %, and the optimal inhibiting dosage is 5.47%. Difference could be detected among the Qingkailing injection samples from different factories and different batches. It is proved that Qingkailing injection could inhibit the metabolic behavior of S. aureus, and microcalorimetry might be applied in the quality assessment of Qingkailing injection.

Comparative study of effects of two bile acid derivatives on Staphylococcus aureus by multiple analytical methods.

The effects of two bile acid derivatives, cholic acid (CA) and deoxycholic acid (DCA) on Staphylococcus aureus (S. aureus) growth were investigated and compared by microcalorimetry coupled with multiple analytical methods. The heat power (HP)-time curves of S. aureus growth affected by CA and DCA were studied by similarity analysis (SA), respectively. Then the quantitative thermo-kinetic parameters obtained from these curves were investigated by the multivariate analysis of variance (MANOVA) and principal component analysis (PCA). By analyzing the two main parameters, growth rate constant k(2) of the second exponential phase and the heat power P(2) of the second highest peak, together with the minimum inhibitory concentration (MIC) values of 10 microg/mL for CA and 20 microg/mL for DCA, it could be concluded that the antibacterial effect of CA was stronger than that of DCA. The existence of alpha-OH at C-7 position of steroid nucleus of bile acid derivatives enhanced the hydrophilicity of compound CA and its inhibitory effect on S. aureus. This study provides a useful method and idea to accurately evaluate the antibacterial effects of bile acid derivatives, which provides some references for screening out new antibacterial agents with high efficacy and low toxicity.

[Study on the best concentration of combination of the effects of the four original constituents of liu-wei di-huang pill on the proliferation and differentiation of rat preadipocyte].

OBJECTIVE: To investigate the effects of four original constituents (Loganin, Morroniside, Peoniflorin, Paeonol) of Liu-wei Di-huang pills on the proliferation and differentiation of rat preadipocyte, and optimize the best concentration by the orthogonal test. METHODS: The rat preadipocytes were cultured, then above four original constituents according to 4 factors and 3 levels to make L9 (3(4)) orthogonal test, and the control was performed at the same time. The proliferation of preadipocytes was determined by MTT method, and the accumulation of cellular lipid was determined by Oil Red O staining, the differences between factors and variances were compared by the value of absorbance. RESULTS: The combination of four original constituents of Liu-wei Di-huang pills could stimulate rat preadipocyte proliferation and inhibited its lipid accumulation. Both the effects were highly significant (P < 0.01) with the combination of A3 B1 C3 D2; Compared with the monosomic group, the effects of the A3 B1 C3 D2 was highly significant (P < 0.01). CONCLUSION: Compared with the monosomic group, the effects of the combination of the four original constituents of Liu-wei Di-huang pills on the proliferation and differentiation of rat preadipocyte were highly significant. This combination consists of Loganin, morroniside with high doses, Paeonol, Peoniflorin with mid-dose and low-dose, respectively. Regnesent the overall regulatory role of synergy and officiencg.

[Cytological and SSR analysis of irradiation-induced progenies of Trititrigia substitution line SN0095].

Pollens of Trititrigia substitution line SN0095 were radiated with 60Co-gamma ray and cytological characters of the progenies M1 and M2 were studied. The results showed that variations with different frequencies and chromosome numbers (2n=41, 2n=40 and 2n=39) were observed in both M1 and M2. Abnormal phenomena, such as univalents, multivalents, chromosome fragments, laggard chromosomes, chromosome bridges and micronucleus, appeared in high frequencies during the meiosis. It suggested that irradiation could promote changes of chromosome number and configuration efficiently, and could lead to rearrangements or translocations between chromosomes. The M2 progenies were examined using BARC159(240), a Thinopyrum intermedium-specific SSR marker of SN0095. Most M2 plants carried the locus BARC159(240), which indicated that the specific segment involving the locus BARC159(240) of Thinopyrum intermedium existed in these plants. However, a band of common wheat YN15 disappeared in a few plants although the locus BARC159(240) existed, which implied that chromosome rearrangements may have occurred in these plants.

[Characterization of Triticum aestivum-Dasypyrum villosum germplasms Shannong05078].

A germplasm line Shannong05078 was developed from the progenies of Am3 x Triticum aestivum-Dasypyrum villosum amphidiploid, the purpose of this study is to ascertain the genome composition of Shannong05078 and then supply wheat breeding with reference. The results showed that Shannong05078 was stable in morphology characters and presenting favorable agronomic traits, it was resistant to powdery mildew and rust. Its protein content was higher, and the HWM-GS of it was 7+8, 5+10. The chromosome number of Shannong05078 was 2n=28, and the chromosome configuration was 2n=14 II at the PMC MI. SSR analysis showed that Shannong05078 contained the whole A and B genomes, and had little genetic materials from D genome. A specific primer of Dasypyrum villosum (phv29) was used to identify the genetic materials form Dasypyrum villosum in Shannong05078, and specific band of Dasypyrum villosum appeared in it. It can be deduced that the genomic composition of Shannong05078 is 2n=AABB=28, and genetic materials of D genome and Dasypyrum villosum were translated into Shannong05078.

[Clinical characteristies and distribution of hepatitis B virus genotype and sub-genotype].

OBJECTIVE: To determine the distribution and virologic characteristics of HBV genotypes, sub-type and possible association with the severity of liver disease. METHODS: 884 patients infected with HBV were enrolled from 8 provinces in China. HBV genotype and sub-type was determined, using PCR-RFLP method. RESULTS: The most common HBV genotypes were B (20.77% ) and C (78.22 % ) but only 1 patient showed genotypes D. We found sub-type Ba in patients with genotype B, C1 and C2 sub-type in patients with genotype C. Genotype C (83.62%) and sub-type C2 (90.32%) were predominant in northern China. Patients with genotype B were much younger than those with genotype C. There was no significant difference between patients with sub-type C1 and C2. There was no significant difference in liver function and serum HBV-DNA load between patients with genotype B and C,or between patients with sub type C1 and C2. However, hepatic inflammation and fibrosis score in patients with genotype B were significantly lower than those with genotype C. CONCLUSION: There were no significantly differences in liver function and HBV-DNA load between patients with genotype B and C, or between patients with sub-type C1 and C2. Hepatic inflammation and fibrosis score in patients with genotype B were significantly lower than those with genotype C. Genotype C/sub-type C2 were preponderance in northern China.

[Analysis of factors influencing GISH used in Triticeae crops' body cells].

Factors influencing genomic in situ hybridization (GISH) used in the body cells of Triticeae crops including Octoploid Tritileymus, Octoploid Trititrigia, Octoploid Tritelytrigia and wheat-Elytrigia intermedium alien disomic addition line were analyzed. The results indicated that ideal GISH effects were based on the high quality chromosome preparations containing more cell mitotic metaphase, better dispersed chromosomes and no foreign matter. The critical factors were suitable ratio of probe DNA to blocker DNA, and the control of elution condition. In addition, some abnormal phenomena, such as chromosome loss, no hybridization signal in alien chromosome, strong or weak hybridization signal, poor or overabundant hybridization signal (high hybridization background), noisy hybridization signal and hybridization spots, were preliminarily analyzed. Proposals and solutions for these problems were further brought forth.

[Cultivation and identification of wheat-Elytrigia intermedium alien disomic addition lines].

6 alien disomic addition lines (line0605; line0607, line0609, line0610, line0611, line0625), which were selected from 59 wheat-Elytrigia intermedium hybridization germ plasm lines, were identified by using the methods of morphology, powdery mildew resistance identifications, cytology and RAPD analysis. Morphogy results showed that they all had the better agronomic characters of their parents; Cytogic results showed that the chromosome configuration at PMC M I was 2n = 22 II; 209 random primers were used for RAPD amplification, result showed 5 of them amplified the specific bands of Elytrigia intermedium parents of 6 disomic addition lines, which could be used as RAPD markers for alien addition chromosomes. Results of powdery mildew resistance showed that line0605 was immune, line0607 was intermediately resistant, line 0610 and line0625 were highly resistant, and line0609 and line0611 were intermediately sensitive.

[Development and molecular cytogenetic identification of 1RS.1BL Translocation lines derived from Triticale x Tritileymus].

In the progenies of octoploid Triticale Jinsong49 crossed with octoploid Tritileymus, a stable germplasm line named Shannong 030-1 was obtained. Its chromosome number was 42, and 21 bivalents could be observed at PMC MI. FISH analysis by using genomic DNAs from S. cereale and genomic DNAs from L. mollis as probes, respectively, showed that Shannong 030-1 contained a pair of translocation chromosome between rye and wheat, and had no genetic materials from L. mollis. C-banding analysis showed that it was translocation lines of 1RS.1BL. Resistance identification showed Shannong 030-1 was immune to powdery mildew.